Human HMGB1 ELISA Kit

Human HMGB1 ELISA Kit

Human HMGB1 ELISA Kit - Information

The ELISA Genie HMGB1 / High mobility group protein B1 ELISA Kit can assay for HMGB1 / High mobility group protein B1 in the following samples: serum, blood, plasma, cell culture supernatant and other related supernatants and tissues.

How our HMGB1 / High mobility group protein B1 ELISA Kits Work?

The ELISA Genie (enzyme-linked immunosorbent assays) assay kits are designed for the quantitative measurement of analytes in a wide variety of samples. As today's scientists demand high quality consistent data for high impact journals, ELISA Genie have developed our range of sensitive, fast and reliable ELISA kit assays to meet and exceed those demands. Our assay kits use a quantitative sandwich ELISA technique and each kit comes with highly specific antibodies pre-coated onto a 96-well microtiter plate.

Human HMGB1 ELISA Kit components	96 Assays	Storage
ELISA Microplate(Dismountable)	8×12 strips	4°C for 6 months
Lyophilized Standard	2	4°C/-20°C
Sample/Standard Dilution Buffer	20ml	4°C
Biotin-labeled Antibody(Concentrated)	120ul	4°C (Protect from light)
Antibody Dilution Buffer	10ml	4°C
HRP-Streptavidin Conjugate(SABC)	120ul	4°C (Protect from light)
SABC Dilution Buffer	10ml	4°C
TMB Substrate	10ml	4°C (Protect from light)
Stop Solution	10ml	4°C
Wash Buffer(25X)	30ml	4°C
Plate Sealer	5	- Kit Protocol: 1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells! 2. Aliquot 0.1ml standard solutions into the standard wells. 3. Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well. 4. Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells. 5. Seal the plate with a cover and incubate at 37 °C for 90 min. 6. Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2. 7. Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall. 8. Seal the plate with a cover and incubate at 37°C for 60 min. 9. Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash. 10. Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min. 11. Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min. 12. Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color. 13. Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. 14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.